Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan 750004, China.
Department of Biochemistry and Molecular Biology, Ningxia Medical University, Yinchuan 750004, China.
J Zhejiang Univ Sci B. 2021 Oct 15;22(10):839-855. doi: 10.1631/jzus.B2000802.
B cell lymphoma 2 () is an important antiapoptotic gene that plays a dual role in the maintenance of the dynamic balance between the survival and death of cancer cells. In our previous study, Bcl-2 was shown to delay the G0/G1 to S phase entry by regulating the mitochondrial metabolic pathways to produce lower levels of adenosine triphosphate (ATP) and reactive oxygen species (ROS). However, the detailed molecular mechanisms or pathways by which Bcl-2 regulates the cell cycle remain unknown. Here, we compared the effects of Bcl-2 overexpression with an empty vector control in the NIH3T3 cell line synchronized by serum starvation, and evaluated the effects using proteomic analysis. The effect of Bcl-2 on cell cycle regulation was detected by monitoring Bcl-2 and p27 expression. The result of subsequent proteomic analysis of Bcl-2 overexpressing cells identified 169 upregulated and 120 downregulated proteins with a 1.5-fold change. These differentially expressed proteins were enriched in a number of signaling pathways predominantly involving the ribosome and oxidative phosphorylation, according to the data of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. These results indicated that Bcl-2 potentially acts at the translation level to influence proteins or enzymes of the respiratory chain or in the ribosome, and thereby regulates the cell cycle. Additionally, differentially expressed proteins involved in oxidative phosphorylation were determined to account for most of the effects of Bcl-2 on the cell cycle mediated by the mitochondrial pathway investigated in our previous study. These results can provide assistance for additional in-depth studies on the regulation of the cell cycle by Bcl-2. The results of the proteomic analysis determined the mechanism of Bcl-2-dependent delay of the cell cycle progression. In summary, the results of this study provide a novel mechanistic basis for identifying the key proteins or pathways for designing and developing precisely targeted cancer drugs.
B 细胞淋巴瘤 2 () 是一种重要的抗凋亡基因，在维持癌细胞存活和死亡的动态平衡中发挥双重作用。在我们之前的研究中，Bcl-2 通过调节线粒体代谢途径来产生较低水平的三磷酸腺苷 (ATP) 和活性氧 (ROS)，从而延迟 G0/G1 期进入 S 期。然而，Bcl-2 调节细胞周期的详细分子机制或途径尚不清楚。在这里，我们比较了 Bcl-2 过表达与空载体对照在血清饥饿同步的 NIH3T3 细胞系中的作用，并通过蛋白质组学分析进行了评估。通过监测 Bcl-2 和 p27 的表达来检测 Bcl-2 对细胞周期调控的影响。随后对 Bcl-2 过表达细胞的蛋白质组学分析结果表明，有 169 个上调蛋白和 120 个下调蛋白的变化倍数为 1.5 倍。根据基因本体论 (GO) 和京都基因与基因组百科全书 (KEGG) 富集分析的数据，这些差异表达蛋白富集在许多信号通路中，主要涉及核糖体和氧化磷酸化。这些结果表明，Bcl-2 可能在翻译水平上作用于呼吸链或核糖体中的蛋白质或酶，从而调节细胞周期。此外，氧化磷酸化涉及的差异表达蛋白被确定为占 Bcl-2 通过我们之前研究的线粒体途径对细胞周期的大部分影响。这些结果可以为进一步深入研究 Bcl-2 对细胞周期的调控提供帮助。蛋白质组学分析的结果确定了 Bcl-2 依赖性延迟细胞周期进程的机制。总之，本研究的结果为确定设计和开发精确靶向癌症药物的关键蛋白或途径提供了新的机制基础。
